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Assessment of Immunogenicity and Protective Efficacy of Zycov-D DNA Vaccine

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Assessment of Immunogenicity and Protective Efficacy of Zycov-D DNA Vaccine bioRxiv preprint doi: https://doi.org/10.1101/2021.02.02.429480; this version posted February 3, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. Assessment of immunogenicity and protective efficacy of ZyCoV-D DNA vaccine candidates in Rhesus macaques against SARS-CoV-2 infection Authors: Pragya D Yadav#1*, Sanjay Kumar#2, Kshitij Agarwal3, Mukul Jain4, Dilip R Patil1, Kapil Maithal5, Basavaraj Mathapati1, Suresh Giri4, Sreelekshmy Mohandas1, Anita Shete1, Gajanan Sapkal1, Deepak Y Patil1, Ayan Dey5, Harish Chandra5, Gururaj Deshpande1, Nivedita Gupta3, Dimpal Nyayanit1, Himanshu Kaushal1, Rima Sahay1, Anuradha Tripathy1, Rajlaxmi Jain1, Abhimanyu Kumar1, Prasad Sarkale1, Shreekant Baradkar1, Chozhavel Rajanathan5, Hari Prasad Raju5, Satish Patel4, Niraj Shah4, Pankaj Dwivedi4, Dharmendra Singh1, Priya Abraham1 #Equal contribution in the study Affiliations: 1Indian Council of Medical Research-National Institute of Virology, Pune, Maharashtra, India Pin-411021 2Dept of Neurosurgery, Command Hospital (Southern Command), Armed Forces Medical College (AFMC), Pune, India Pin- 411040 3Indian Council of Medical Research, Ramalingaswami Bhawan, P.O. Box No. 4911, Ansari Nagar, New Delhi, India Pin-110029 4Zydus Research Centre, Cadila Healthcare Limited, Ahmedabad, Gujarat, India Pin-382481 5Vaccine Technology Centre, Cadila Healthcare Limited, Ahmedabad, Gujarat, India Pin-382481 Corresponding author*: Dr. Pragya D Yadav, Scientist ‘E’ and Group Leader, 1 bioRxiv preprint doi: https://doi.org/10.1101/2021.02.02.429480; this version posted February 3, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. Maximum Containment Facility, Indian Council of Medical Research- National Institute of Virology, Sus Road, Pashan, Pune–411 021, India. Phone: +9120-26006111, Fax No. 91-20-26122669 Email: [email protected] Abstract Vaccines remain the key protective measure to achieve herd immunity to control the disease burden and stop COVID-19 pandemic. We have developed and assessed the immunogenicity and protective efficacy of two formulations (1mg and 2mg) of ZyCoV-D (a plasmid DNA based vaccine candidates) administered through Needle Free Injection System (NFIS) and syringe- needle (intradermal) in rhesus macaques with three dose vaccine regimens. The vaccine candidate 2mg dose administered using Needle Free Injection System (NFIS) elicited a significant immune response with development of SARS-CoV-2 S1 spike region specific IgG and neutralizing antibody (NAb) titers during the immunization phase and significant enhancement in the levels after the virus challenge. In 2 mg NFIS group the IgG and NAb titers were maintained and showed gradual rise during the immunization period (15 weeks) and till 2 weeks after the virus challenge. It also conferred better protection to macaques evident by the viral clearance from nasal swab, throat swab and bronchoalveolar lavage fluid specimens in comparison with macaques from other immunized groups. In contrast, the animals from placebo group developed high levels of viremia and lung disease following the virus challenge. Besides this, the vaccine candidate also induced increase lymphocyte proliferation and cytokines response (IL-6, IL-5).The administration of the vaccine candidate with NFIS generated a better immunogenicity response in comparison to syringe-needle (intradermal route). The study bioRxiv preprint doi: https://doi.org/10.1101/2021.02.02.429480; this version posted February 3, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. demonstrated immunogenicity and protective efficacy of the vaccine candidate, ZyCoV-D in rhesus macaques. Keywords: SARS-CoV-2; DNA vaccine; Nonhuman primate; viremia; Neutralizing antibodies INTRODUCTION The ongoing pandemic of Coronavirus disease-19 (COVID-19) has affected countries across the globe and millions of people have been impacted leading to loss of life, economy and productivity. This has led to development of various intervention strategies including development of vaccine for prophylactic use, monoclonal antibodies and other biologics for therapeutic application.Two mRNA-based vaccines (1) have been granted emergency use authorization (EUA) in UK, USA, Canada, European Union and many other countries across the world. Two adenovirus based non-replicating viral vaccine (Oxford and Sputnik) has been granted EUA by UK, India and Russia. However, the need for additional vaccine candidates still persists to so that vaccines would be available to a larger population across the world. Studies have utilized various vaccine development platforms such as inactivated whole virion, live- attenuated, nucleic acid-based (RNA, DNA), replicating /non-replicating viral vector-based, protein subunit and numerous other platforms (2). The response of the global scientific community has resulted in development of over 233 vaccine candidates until December 29 2020. Of these, 172 candidates are in pre-clinical stage and 61 candidates have progressed to clinical trials (3). One of thesafe and efficacious next generation vaccines is the DNA vaccine which involves direct administration of plasmid DNA encoding for immunogenic antigen component of the pathogen (4). DNA vaccines are composed of bacterial plasmids with a gene encoding for the protein of interest and transcription promoter and terminator. Routes of administration of DNA vaccines can be topical application, parenteral or cytofectin-mediated method. The plasmid gains bioRxiv preprint doi: https://doi.org/10.1101/2021.02.02.429480; this version posted February 3, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. entry in multiple cells such as myocytes, keratinocytes, and antigen-presenting cells (APCs) where it enters in the nucleus as an episome; without getting integrated into the host cell DNA. Using the host cell’s transcription and translation machinery, the inserted gene gets translatedinto antigen. Protein expressed by plasmid-transfected cells is likely to be expressed within the cell and folded in its native configuration. The antigen is recognized by antigen presenting cells (APCs) and further induces antibodies and cellular response. DNA vaccines are free of any infectious agent, are low cost, fairly safe and relatively easy for large-scale manufacturing (5). They induce strong and long-lasting humoral and cell-mediated immune response (6). DNA vaccines are heat-stable that makes the storage and transport of these vaccine easy (7). Preclinical efficacy of DNA vaccines has been demonstrated in animal models against SARS (8), MERS (9), influenza virus, human immunodeficiency virus, rabies virus, hepatitis B virus, lymphocytic choriomeningitis virus, malarial parasitesand mycoplasma (10, 11). Studies have reported higher immune response of DNA vaccine by their administration using spring-powered and needle-free devices in animals such as mice, chickens, pigs and non-human primates (NHP) (12-17). The delivery procedure through skin is less painful and it produces higher immunogenicity because of abundance of antigen-presenting cells and dermal lymphatic vessels in skin (18). Currently, there are twenty-four DNA-based vaccine candidates developed by different pharmaceutical/biotech companies. Sixteen of themare in pre-clinical stage, while eight have progressed toclinical trials (3). ZyCoV-D DNA vaccine candidate developed by Cadila Healthcare Limited has recently completed phase I/II clinical trial (CTRI/2020/07/026352). Here, we report the development and evaluation of the immunogenicity and protective efficacy of ZyCoV-D DNA vaccine candidates in rhesus macaques against SARS-CoV-2 infection. bioRxiv preprint doi: https://doi.org/10.1101/2021.02.02.429480; this version posted February 3, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. RESULTS Anti-SARS-CoV-2 IgG response We evaluated anti-SARS-CoV-2 Immunoglobulin-G (IgG) antibody response against the spike proteinS1 region from the macaque serum samples during the immunization phase (0, 28, 42, 56,70, 84 and 103 days) and SARS-CoV-2 challenge [0, 1, 3, 5, 7, 11 and 15 days post infection (DPI)]. During the immunization phase, animals of group II (1 mg /NFIS) and group IV (2 mg /NFIS) elicited IgG (S1) response starting from day 42 and a gradual rise till day 103 (Fig. 1. Aand C). Animals of group III (2 mg /syringe needle) had detectable IgG (S1) levels starting at day 28, but the response was not significanttill day 103. Group I animals (1 mg /syringe needle) and group V (placebo) did not show IgG (S1) response throughout the period of immunization phase (Fig. 1. A and C). After the
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